scholarly journals Localization (and profiles) of tyrosinephosphorylated proteins in female reproductive organs of adult rats

2020 ◽  
Vol 47 (3) ◽  
pp. 180-185
Author(s):  
Sudtida Bunsueb ◽  
Nareelak Tangsrisakda ◽  
Alexander T.H. Wu ◽  
Sitthichai Iamsaard
Keyword(s):  
Stromal Cells ◽  
Expression Patterns ◽  
Sodium Dodecyl ◽  
Reproductive Organs ◽  
Uterine Wall ◽  
Apical Surface ◽  
Sprague Dawley ◽  
Adult Rats ◽  
Sprague Dawley Rats ◽  
Estrus Phase

Objective: Tyrosine phosphorylation is an essential process in many biological systems, including the male reproductive system. The presence of tyrosine-phosphorylated (TyrPho) proteins has been well documented in male reproductive organs, but research in fertile females is still limited.Methods: The ovary, oviduct, and uterus of adult female Sprague-Dawley rats in the estrus phase were used to localize TyrPho proteins using an immunohistochemical technique. These proteins were separated and their expression patterns were examined by sodium dodecyl sulfatepolyacrylamide gel electrophoresis and Western blot analysis, respectively.Results: TyrPho proteins were localized in the cytoplasm of the oocyte except the antral fluid; in the granulosa cells, theca cells, and stromal cells of the ovary; at the apical surface of oviductal epithelial cells; and in the basal epithelium and submucosa of the uterine wall. Moreover, we found that 72-, 43-, and 28-kDa TyrPho proteins were localized in the ovary, while 170-, 55-, and 43-kDa proteins were localized in the oviduct. In the uterus, we detected four major bands, corresponding to 61-, 55-, 54-, and 43-kDa TyrPho proteins.Conclusion: Given that these TyrPho proteins were found in major reproductive organs in the estrus phase, these proteins may play important roles in female fertility.

scholarly journals Chronic stress increases the tyrosine phosphorylation in female reproductive organs: An experimental study

2021 ◽  
Author(s):  
Sudtida Bunsueb ◽  
Natthapol Lapyuneyong ◽  
Saranya Tongpan ◽  
Supatcharee Arun ◽  
Sitthichai Iamsaard
Keyword(s):  
Experimental Study ◽  
Tyrosine Phosphorylation ◽  
Chronic Stress ◽  
Cold Water ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Reproductive Organs ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Sds Page

Background: Changes in tyrosine-phosphorylated (TyrPho) protein expressions have demonstrated stress in males. In females, chronic stress (CS) is a major cause of infertility, especially anovulation. However, the tyrosine phosphorylation in the female reproductive system under stress conditions has never been reported. Objective: To investigate the alteration of TyrPho protein expression in ovary, oviduct, and uterus of CS rats. Materials and Methods: In this experimental study, 16 female Sprague-Dawley rats (5 wk: 220-250 gr) were divided into control and CS groups (n = 8/group). Every day, the CS animals were immobilized within a restraint cage and individually forced to swim in cold water for 60 consecutive days. Following the stress induction, the ovary, oviduct, and uterus of all rats were observed for their morphologies. The total protein profiles of all tissues were revealed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) before detecting TyrPho proteins using western blot. Intensity analysis was used to compare the expression of proteins between groups. Results: The results showed that the morphology and weights of ovary and oviduct in the CS group were not different from control. In contrast, the CS significantly increased the uterine weight as compared to control. Moreover, the expressions of TyrPho proteins in the ovary (72, 43, and 28 kDas), oviduct (170, 55, and 43 kDas), and uterus (55, 54, and 43 kDas) were increased in CS group as compared to those of control. Conclusion: The increased expressions of TyrPho proteins in ovary, oviduct, and uterus could be potential markers used to explain some mechanisms of female infertility caused by chronic stress. Key words: Ovary, Oviduct, Uterus, Phosphorylation.

scholarly journals Uterine Paramesonephric Cysts in Sprague-Dawley Rats from National Toxicology Program Studies

2018 ◽  
Vol 46 (4) ◽  
pp. 421-430
Author(s):  
Daven N. Jackson-Humbles ◽  
John Curtis Seely ◽  
Ronald A. Herbert ◽  
David E. Malarkey ◽  
Barry S. McIntyre ◽  
...  
Keyword(s):  
Estrogen Receptor Α ◽  
Uterine Wall ◽  
Sprague Dawley ◽  
Cytokeratin 18 ◽  
Adult Rats ◽  
National Toxicology Program ◽  
Sd Rats ◽  
Sprague Dawley Rats ◽  
Gata3 Expression ◽  
Flocculent Material

Congenital uterine wall cysts arising from paramesonephric (Müllerian) and mesonephric (Wolffian) ducts are typically incidental findings in most species. We used immunohistochemistry to characterize and determine the origin of uterine cysts in Sprague-Dawley (SD) rats from multigeneration studies conducted by the National Toxicology Program. Subserosal uterine cysts were observed in 20 of the 2,400 SD rats evaluated in five studies, and 10 cysts were characterized for this study. Single cysts were unilocular, fluid-filled, and occurred throughout the uterus. Microscopically, all cysts had a well-developed smooth muscle wall, lined by flattened to cuboidal, sometimes ciliated, epithelium that stained intensely positive for cytokeratin 18 and paired box protein 8 (PAX8). Most cyst epithelia displayed weak to moderate positivity for progesterone receptor (PR) and/or estrogen receptor α (ER-α), as well as were negative for GATA binding protein 3 (GATA3). Cyst lumens contained basophilic flocculent material. The cysts appeared to be developmental anomalies arising from paramesonephric tissue based on positive PAX8 and ER-α and/or PR staining. Additionally, 70% of the cysts lacked GATA3 expression. Taken together, the subserosal uterine cysts observed in adult rats in these studies most likely arose from the paramesonephric duct.

Sepsis induces changes in the expression and distribution of Toll-like receptor 4 in the rat kidney

2006 ◽  
Vol 290 (5) ◽  
pp. F1034-F1043 ◽  
Author(s):  
Tarek M. El-Achkar ◽  
Xiaoping Huang ◽  
Zoya Plotkin ◽  
Ruben M. Sandoval ◽  
Georges J. Rhodes ◽  
...  
Keyword(s):  
Rat Kidney ◽  
Expression Patterns ◽  
Cecal Ligation ◽  
Proximal Tubules ◽  
Microbial Pathogens ◽  
Sprague Dawley ◽  
Renal Vasculature ◽  
Two Photon Microscopy ◽  
Sprague Dawley Rats ◽  
Cellular Signal

Toll-like receptors (TLRs) are now recognized as the major receptors for microbial pathogens on cells of the innate immune system. Recently, TLRs were also identified in many organs including the kidney. However, the cellular distribution and role of these renal TLRs remain largely unknown. In this paper, we investigated the expression of TLR4 in a cecal ligation and puncture (CLP) model of sepsis in Sprague-Dawley rats utilizing fluorescence microscopy. In sham animals, TLR4 was expressed predominantly in Tamm-Horsfall protein (THP)-positive tubules. In CLP animals, TLR4 expression increased markedly in all tubules (proximal and distal), glomeruli, and the renal vasculature. The staining showed a strong apical distribution in all tubules. A moderately less intense cellular signal colocalized partially with the Golgi apparatus. In addition, kidneys from septic rats showed increased expression of CD14 and THP. They each colocalized strongly with TLR4, albeit in different tubular segments. We also imaged the kidneys of live septic animals with two-photon microscopy after fluorescent lipopolysaccharide (LPS) injection. Within 10 min, LPS was seen at the brush border of some proximal tubules. Within 60 min, LPS was fully cytoplasmic in proximal tubules. Conversely, distal tubules showed no LPS uptake. We conclude that TLR4, CD14, and THP have specific renal cellular and tubular expression patterns that are markedly affected by sepsis. Systemic endotoxin can freely access the tubular and cellular sites where these proteins are present. Therefore, locally expressed TLRs and other interacting proteins could potentially modulate the renal response to systemic sepsis.

scholarly journals Effects of adolescent caffeine consumption on cocaine self-administration and reinstatement of cocaine seeking

2018 ◽  
Vol 33 (1) ◽  
pp. 132-144
Author(s):  
Tracey A Larson ◽  
Casey E O’Neill ◽  
Michaela P Palumbo ◽  
Ryan K Bachtell
Keyword(s):  
Dose Response ◽  
Extinction Training ◽  
Schedules Of Reinforcement ◽  
Sprague Dawley ◽  
Self Administration ◽  
Caffeine Consumption ◽  
Adult Rats ◽  
Sprague Dawley Rats ◽  
Cocaine Seeking ◽  
Administration Procedure

Background: Caffeine consumption by children and adolescents has risen dramatically in recent years, yet the lasting effects of caffeine consumption during adolescence remain poorly understood. Aim: These experiments explore the effects of adolescent caffeine consumption on cocaine self-administration and seeking using a rodent model. Methods: Sprague-Dawley rats consumed caffeine for 28 days during the adolescent period. Following the caffeine consumption period, the caffeine solution was replaced with water for the remainder of the experiment. Age-matched control rats received water for the duration of the study. Behavioral testing in a cocaine self-administration procedure occurred during adulthood (postnatal days 62–82) to evaluate how adolescent caffeine exposure influenced the reinforcing properties of cocaine. Cocaine seeking was also tested during extinction training and reinstatement tests following cocaine self-administration. Results: Adolescent caffeine consumption increased the acquisition of cocaine self-administration and increased performance on different schedules of reinforcement. Consumption of caffeine in adult rats did not produce similar enhancements in cocaine self-administration. Adolescent caffeine consumption also produced an upward shift in the U-shaped dose response curve on cocaine self-administration maintained on a within-session dose-response procedure. Adolescent caffeine consumption had no effect on cocaine seeking during extinction training or reinstatement of cocaine seeking by cues or cocaine. Conclusions: These findings suggest that caffeine consumption during adolescence may enhance the reinforcing properties of cocaine, leading to enhanced acquisition that may contribute to increased addiction vulnerability.

scholarly journals Protective effects of dietary fibre against manganese-induced neurobehavioral aberrations in rats

2012 ◽  
Vol 63 (3) ◽  
pp. 263-270 ◽  
Author(s):  
Xiu-Quan Shi ◽  
Wei Yan ◽  
Ke-Yue Wang ◽  
Qi-Yuan Fan ◽  
Yan Zou
Keyword(s):  
Cerebral Cortex ◽  
Animal Feed ◽  
Oral Exposure ◽  
Protective Effects ◽  
Sprague Dawley ◽  
Adult Rats ◽  
Neurobehavioral Performance ◽  
Sprague Dawley Rats ◽  
Applied Dose ◽  
Mn Concentrations

We tested the hypothesis that dietary fi bre (DF) has protective effects against manganese (Mn)-induced neurotoxicity. Forty-eight one-month old Sprague-Dawley rats were randomly divided into six groups: control, 16 % DF, Mn (50 mg kg-1 body weight), Mn+ 4 % DF, Mn+ 8 % DF, and Mn+ 16 % DF. After oral administration of Mn (as MnCl2) by intragastric tube during one month, we determined Mn concentrations in the blood, liver, cerebral cortex, and stool and tested neurobehavioral functions. Administration of Mn was associated with increased Mn concentration in the blood, liver, and cerebral cortex and increased Mn excretion in the stool. Aberrations in neurobehavioral performance included increases in escape latency and number of errors and decrease in step-down latency. Irrespective of the applied dose, the addition of DF in forage decreased tissue Mn concentrations and increased Mn excretion rate in the stool by 20 % to 35 %. All neurobehavioral aberrations were also improved. Our fi ndings show that oral exposure to Mn may cause neurobehavioral abnormalities in adult rats that could be effi ciently alleviated by concomitant supplementation of DF in animal feed.

scholarly journals Superoxide Dismutase Expression in the Ovaries of Periodontitis Animal Models Induced by Porphyromonas gingivalis and Treated with Cassava Leaves Extract

2021 ◽  
Vol 28 ◽  
pp. 40-46
Author(s):  
Agustin Wulan Suci-Dharmayanti ◽  
Ajeng Nurwahyuningtyas Anjani ◽  
Ari Tri Wanodyo Handayani ◽  
Zahreni Hamzah ◽  
Zahara Meilawaty ◽  
...  
Keyword(s):  
Animal Models ◽  
Porphyromonas Gingivalis ◽  
Reproductive Organs ◽  
Sprague Dawley ◽  
Sex Steroid Hormone ◽  
Low Intensity ◽  
Fertility Status ◽  
Cassava Leaves ◽  
Sprague Dawley Rats ◽  
Female Reproductive Organs

The ovaries represent the female reproductive organs that determine the women's fertility status and their systemic and oral health, correlating to sex steroid hormone alteration. This study aimed to investigate the effect of cassava leaves extract treatment to SOD expression in the animal model-ovaries after Porphyromonas gingivalis injection. 15 female Sprague Dawley rats were used and divided into five groups: (1) control without cassava leaves extract treatment (C); (2) P. gingivalis without cassava leaves extract treatment (T1); (3) P. gingivalis and cassava leaves extract (T2); (4) P. gingivalis and vitamin C (T3); and (5) P. gingivalis and metronidazole (T4). Animal were euthanised at day seven after the initial treatment to collect ovaries. The ovaries sections were immunohistochemically stained to quantify SOD expression using light microscope while the Image J software was used to quantify the SOD expression. The results showed that all of the follicle types had the same intensity of SOD expression. Most of the follicles exhibited low intensity of SOD expression, except for atretic follicles. In conclusion, P. gingivalis and cassava leaves extract influenced SOD expression in the ovaries of animal models, which increased the SOD expression.

Kinetic impairment of nitrogen and muscle glutamine metabolisms in old glucocorticoid-treated rats

1999 ◽  
Vol 276 (3) ◽  
pp. E558-E564 ◽  
Author(s):  
Regine Minet-Quinard ◽  
Christophe Moinard ◽  
Françoise Villie ◽  
Stephane Walrand ◽  
Marie-Paule Vasson ◽  
...  
Keyword(s):  
Control Group ◽  
Synthetase Activity ◽  
Aged Rats ◽  
Sprague Dawley ◽  
Adult Rats ◽  
Glucocorticoid Treatment ◽  
Sprague Dawley Rats ◽  
End Of Treatment ◽  
Catabolic State ◽  
Old Rats

Aged rats are more sensitive to injury, possibly through an impairment of nitrogen and glutamine (Gln) metabolisms mediated by glucocorticoids. We studied the metabolic kinetic response of adult and old rats during glucocorticoid treatment. The male Sprague-Dawley rats were 24 or 3 mo old. Both adult and old rats were divided into 7 groups. Groups labeled G3, G5, and G7 received, by intraperitoneal injection, 1.50 mg/kg of dexamethasone (Dex) for 3, 5, and 7 days, respectively. Groups labeled G3PF, G5PF, and G7PF were pair fed to the G3, G5, or G7 groups and were injected with an isovolumic solution of NaCl. One control group comprised healthy rats fed ad libitum. The response to aggression induced specifically by Dex (i.e., allowing for variations in pair-fed controls) appeared later in the aged rats (decrease in nitrogen balance from day 1 in adults but only from day 4 in old rats). The adult rats rapidly adapted to Dex treatment, whereas the catabolic state worsened until the end of treatment in the old rats. Gln homeostasis was not maintained in the aged rats; despite an early increase in muscular Gln synthetase activity, the Gln pool was depleted. These results suggest a kinetic impairment of both nitrogen and muscle Gln metabolisms in response to Dex with aging.

scholarly journals Synergistic toxicity in an in vivo model of neurodegeneration through the co-expression of human TDP-43M337V and tauT175D protein

2019 ◽  
Vol 7 (1) ◽  
Author(s):  
Alexander J. Moszczynski ◽  
Madeline Harvey ◽  
Niveen Fulcher ◽  
Cleusa de Oliveira ◽  
Patrick McCunn ◽  
...  
Keyword(s):  
Tau Protein ◽  
Sensorimotor Gating ◽  
In Vivo Model ◽  
Motor Deficits ◽  
Sprague Dawley ◽  
Tau Pathology ◽  
Adult Rats ◽  
Sprague Dawley Rats ◽  
Somatic Gene Transfer

Abstract Although it has been suggested that the co-expression of multiple pathological proteins associated with neurodegeneration may act synergistically to induce more widespread neuropathology, experimental evidence of this is sparse. We have previously shown that the expression of Thr175Asp-tau (tauT175D) using somatic gene transfer with a stereotaxically-injected recombinant adeno-associated virus (rAAV9) vector induces tau pathology in rat hippocampus. In this study, we have examined whether the co-expression of human tauT175D with mutant human TDP-43 (TDP-43M337V) will act synergistically. Transgenic female Sprague-Dawley rats that inducibly express mutant human TDP-43M337V using the choline acetyltransferase (ChAT) tetracycline response element (TRE) driver with activity modulating tetracycline-controlled transactivator (tTA) were utilized in these studies. Adult rats were injected with GFP-tagged tau protein constructs in a rAAV9 vector through bilateral stereotaxic injection into the hippocampus. Injected tau constructs were: wild-type GFP-tagged 2N4R human tau (tauWT; n = 8), GFP-tagged tauT175D 2N4R human tau (tauT175D, pseudophosphorylated, toxic variant, n = 8), and GFP (control, n = 8). Six months post-injection, mutant TDP-43M337V expression was induced for 30 days. Behaviour testing identified motor deficits within 3 weeks after TDP-43 expression irrespective of tau expression, though social behaviour and sensorimotor gating remained unchanged. Increased tau pathology was observed in the hippocampus of both tauWT and tauT175D expressing rats and tauT175D pathology was increased in the presence of cholinergic neuronal expression of human TDP-43M337V. These data indicate that co-expression of pathological TDP-43 and tau protein exacerbate the pathology associated with either individual protein.

scholarly journals Cerebral cortex does not modulate “regulated” decrease in core temperature during hypoxemia in rats

1998 ◽  
Vol 274 (4) ◽  
pp. R1158-R1161
Author(s):  
Evvi-Lynn M. Rollins ◽  
James E. Fewell
Keyword(s):  
Cerebral Cortex ◽  
Core Temperature ◽  
Cortical Spreading Depression ◽  
Spreading Depression ◽  
Temperature Response ◽  
Sprague Dawley ◽  
Adult Rats ◽  
The Core ◽  
Sprague Dawley Rats ◽  
Before And After

In newborns and adults of a number of species including humans, exposure to acute hypoxemia produces a “regulated” decease in core temperature, the mechanism of which is unknown. Considering that various cortical areas participate in autonomic regulation including thermoregulation, the present experiments were carried out to test the hypothesis that the cerebral cortex plays a role in modulating the regulated decrease in core temperature during acute hypoxemia. This hypothesis was tested by determining the core temperature response to acute hypoxemia in chronically instrumented adult Sprague-Dawley rats before and after cortical spreading depression (i.e., functional decortication) was produced by the local application of potassium chloride to the dura overlying the cerebral hemispheres. There was no effect of cortical spreading depression on baseline core temperature. Core temperature decreased during acute hypoxemia in a similar fashion when the cerebral cortex was intact as well as during functional decortication. Thus our data do not support the hypothesis that the cerebral cortex modulates the regulated decrease in core temperature that occurs in adult rats during acute hypoxemia.

Characterization of vasoactive intestinal peptide receptors on rat alveolar macrophages

1994 ◽  
Vol 267 (3) ◽  
pp. L256-L262 ◽  
Author(s):  
H. Sakakibara ◽  
K. Shima ◽  
S. I. Said
Keyword(s):  
Vasoactive Intestinal Peptide ◽  
Alveolar Macrophages ◽  
Inflammatory Responses ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
High Capacity ◽  
Sprague Dawley ◽  
Major Band ◽  
High Affinity ◽  
Sprague Dawley Rats

In view of the evidence that vasoactive intestinal peptide (VIP) may modulate acute inflammatory injury in the lung, we investigated the presence and characteristics of VIP receptors on alveolar macrophages (AMs). We examined the binding of monoiodinated [Tyr(125I)10]-labeled VIP (125I-VIP) to rat AMs (> 96% pure), obtained from Sprague-Dawley rats by bronchoalveolar lavage (BAL). At 23 degrees C, the interaction of 125I-VIP with AMs was rapid, reversible, saturable, and linearly proportional to the number of cells. At equilibrium, the binding was competitively inhibited by 10(-11)-10(-6) M of native peptide [half-maximal inhibition (IC50) = 0.53 +/- 0.34 nM, n = 8], with evidence for two classes of binding sites: one with a high affinity (Kd = 0.20 +/- 0.09 nM) and a low capacity (1,190 +/- 640 sites/cell) and another with a low affinity (Kd = 43.2 +/- 13.8 nM) and a high capacity (51,700 +/- 14,000 sites/cell). VIP-related peptides inhibited the binding with the order of potency: VIP > peptide histidine isoleucine > helodermin >> secretin; glucagon was ineffective. In the presence of 3-isobutyl-1-methylxanthine, VIP dose dependently stimulated adenosine 3',5'-cyclic monophosphate accumulation in intact AMs, with maximal stimulation (6.3 times basal level) at 1 nM, and half-maximal accumulation at 0.23 +/- 0.11 nM VIP (Kd for high-affinity sites). For determination of the mass of the VIP receptor, 125I-VIP was covalently bound to AMs with the cross-linking agent dithiobis succinimidyl propionate. Autoradiographic studies after sodium dodecyl sulfate/polyacrylamide gel electrophoresis of solubilized affinity-labeled cells revealed a single major band of M(r) 76,400. We conclude that VIP binds to specific receptors on rat AMs that are coupled to adenylate cyclase, through which VIP may modulate inflammatory responses within the lung.

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